Short-Term Supplementation of Sodium Nitrate vs. Sodium Chloride Increases Homoarginine Synthesis in Young Men Independent of Exercise.

The aim of the study was to investigate the effects of short-term oral administration of inorganic nitrate (NaNO3; n = 8) or placebo (NaCl; n = 9) (each 0.1 mmol/kg body weight/d for 9 days) on plasma amino acids, creatinine, and oxidative stress in healthy young men. At baseline, the plasma concentrations of amino acids did not differ between the groups. At the end of the study, the plasma concentrations of homoarginine (hArg; by 24%, p = 0.0001), citrulline and ornithine (Cit/Orn; by 16%, p = 0.015), and glutamine/glutamate (Gln/Glu; by 6%, p = 0.0003) were higher in the NaNO3 group compared to the NaCl group. The plasma concentrations of sarcosine (Sarc; by 28%, p < 0.0001), tyrosine (by 14%, p = 0.0051), phenylalanine (by 8%, p = 0.0026), and tryptophan (by 8%, p = 0.0047) were lower in the NaNO3 group compared to the NaCl group. These results suggest that nitrate administration affects amino-acid metabolism. The arginine/glycine amidinotransferase (AGAT) catalyzes two reactions: (1) the formation of l-homoarginine (hArg) and l-ornithine (Orn) from l-arginine (Arg) and l-lysine (Lys): Arg + Lys <−> hArg + Orn, with equilibrium constant Kharg; (2) the formation of guanidinoacetate (GAA) and Orn from Arg and glycine (Gly): Arg + Gly <−> GAA + Orn, with equilibrium constant Kgaa. The plasma Kgaa/KhArg ratio was lower in the NaNO3 group compared to the NaCl group (1.57 vs. 2.02, p = 0.0034). Our study suggests that supplementation of inorganic nitrate increases the AGAT-catalyzed synthesis of hArg and decreases the N-methyltransferase-catalyzed synthesis of GAA, the precursor of creatine. To our knowledge, this is the first study to demonstrate elevation of hArg synthesis by inorganic nitrate supplementation. Remarkably, an increase of 24% corresponds to the synthesis capacity of one kidney in healthy humans. Differences in the association between plasma concentrations of amino acids in the NaNO3 and NaCl groups suggest changes in amino-acid homeostasis. Plasma concentrations of the oxidative stress marker malondialdehyde (MDA) did not change after supplementation of NaNO3 or NaCl over the whole exercise time range. Plasma nitrite concentration turned out to be a more discriminant marker of NaNO3 ingestion than plasma nitrate (area under the receiver operating characteristic curve: 0.951 vs. 0.866, p < 0.0001 each).

Everyday Pedelec Use and Its Effect on Meeting Physical Activity Guidelines.

Pedelecs (e-bikes with electrical support up to 25 km·h-1) are important in active transportation. Yet, little is known about physiological responses during their everyday use. We compared daily pedelec (P) and bicycle (B) use to determine if pedelecs are a suitable tool to enhance physical activity. In 101 employees, cycling duration and intensity, heart rate (HR) during P and B were recorded via a smartphone app. Each recording period was a randomized crossover design and lasted two weeks. The ride quantity was higher in P compared to B (5.3 ± 4.3 vs. 3.2 ± 4.0 rides·wk-1; p < 0.001) resulting in a higher total cycling time per week for P (174 ± 146 min·wk-1) compared to B (99 ± 109 min·wk-1; p < 0.001). The mean HR during P was lower than B (109 ± 14 vs. 118 ± 17 bpm; p < 0.001). The perceived exertion was lower in P (11.7 ± 1.8 vs. 12.8 ± 2.1 in B; p < 0.001). The weekly energy expenditure was higher during P than B (717 ± 652 vs. 486 ± 557 metabolic equivalents of the task [MET]·min·wk-1; p < 0.01). Due to a sufficient HR increase in P, pedelecs offer a more active form of transportation to enhance physical activity.

Long-term effects of NO3- on the relationship between oxygen uptake and power after three weeks of supplemented HIHVT.

The aim of this study was to investigate the later effects of daily NO3- supplementation over 3 wk of training on the relationship between O2 uptake and power at different intensities with an incremental test (IT), a double-wingate test (WT), and an endurance capacity test at 80% Wmax (ECT) before and after the supplementation period. Seventeen male recreational athletes participated in this double-blind placebo (PL)-controlled study. Subjects participated in a 3-wk intermittent high-intensity, high-volume training period with 45 intervals of Wmax - 10 W and an active recovery period of 10 W in between with dietary NO3- (NaNO3) or placebo supplementation (NaCl) (both 8.5 mg·kg-1·day-1) on a cycle ergometer. During a training session, plasma [ NO3- ] ( P < 0.001) and plasma [ NO2- ] ( P < 0.01) were higher in nitrate (N), whereas in pre- and posttests mean plasma [ NO3- ] and [ NO2- ] were not different between groups. In the WT [48 h after cessation of supplementation (C)], the ratio between V̇o2 and power decreased in N ( P < 0.01) with no changes in PL. Endurance capacity (4-5 days after C) similarly increased in both groups ( P < 0.01). However, the total oxygen consumption decreased by 5% ( P < 0.01) in N, with no change in PL. The slope of V̇o2·W-1 in IT (5-7 days after C) decreased in N ( P < 0.01), whereas no changes were found in PL. During low- and moderate-intensity workloads, no changes and differences in V̇o2 could be detected. We conclude that nitrate supplementation causes a sustaining reduction of the oxygen cost per watt during exercise with a large recruitment of type II muscle fibers without affecting endurance capacity. NEW & NOTEWORTHY Because most studies focused on the acute effects of NO3- supplementation on exercise performance during a supplementation period, the sustainability of the effects of the NO3- supplementation remain unknown. We followed the development of V̇o2/W at different intensities during the first week after cessation of daily NO3- supplementation over 3 wk. The results indicate that NO3- supplementation has a long-term effect for at least 7 days after cessation during heavy all-out workloads without affecting endurance capacity.

Simultaneous GC-ECNICI-MS measurement of nitrite, nitrate and creatinine in human urine and plasma in clinical settings.

Creatinine in urine is a useful biochemical parameter to correct the urinary excretion rate of endogenous and exogenous substances. Nitrite (ONO-) and nitrate (ONO2-) are metabolites of nitric oxide (NO), a signalling molecule with multiple biological functions. Under certain and standardized conditions, the concentration of nitrate in the urine is a suitable measure of whole body NO synthesis. The urinary nitrate-to-nitrite molar ratio (UNOxR) may indicate nitrite-dependent renal carbonic anhydrase (CA) activity. In clinical studies, urine is commonly collected by spontaneous micturition. In those cases the nitrate and nitrite excretion must be corrected for creatinine excretion. Pentafluorobenzyl (PFB) bromide (PFB-Br) is a useful derivatization reagent of numerous inorganic and organic compounds, including urinary nitrite, nitrate and creatinine, for highly sensitive and specific quantitation by GC-MS. Here, we report on the simultaneous PFB-Br derivatization (60min, 50°C) of ONO-, O15NO-, ONO2-, O15NO2-, creatinine (do-Crea) and [methylo-2H3]creatinine (d3-Crea) in acetonic dilutions of native human urine and plasma samples (4:1, v/v) and their simultaneous quantification by GC-MS as PFBNO2, PFB15NO2, PFBONO2, PFBO15NO2, do-Crea-PFB and d3-Crea-PFB, respectively. Electron capture negative-ion chemical ionization (ECNICI) of these derivatives generates anions due to [M-PFB]-, i.e., the starting analytes. Quantification is performed by selected-ion monitoring (SIM) of m/z 46 (ONO-), m/z 47 (O15NO-), m/z 62 (ONO2-), m/z 63 (O15NO2-), m/z 112 (do-Crea), and m/z 115 (d3-Crea). Retention times were 2.97min for PFB-ONO2/PFB-O15NO2, 3.1min for PFB-NO2/PFB-15NO2, and 6.7min for do-Crea-PFB/d3-Crea-PFB. We used this method to investigate the effects of long-term oral NaNO3 or NaCl (serving as placebo) supplementation (each 0.1mmol/kg body weight per day for 3 weeks) on creatinine excretion and UNOxR in 17 healthy young men. Compared to NaCl (n=8), NaNO3 (n=9) supplementation increased UNOxR (1709±355 vs. 369±77, P<0.05). Creatinine excretion did not differ between the groups (6.67±1.34mM vs. 5.72±1.27mM, P=0.57). The method is also applicable to human plasma. In 78 adults patients newly diagnosed for cerebrovascular disease (CVD), there was a close correlation (r=0.9833) between the creatinine concentrations measured in plasma by GC-ECNICI-MS and those measured in serum by an enzymatic assay. Creatinine-corrected plasma nitrate and nitrite concentrations (P=0.035 and P=0.004, respectively) but not their concentrations (P=0.68 and P=0.40, respectively) differ between male (n=54) and female (n=24) CVD patients. No such differences were found between preterm newborn boys (n=25) and girls (n=22). Like in urine, circulating creatinine may be useful to correct for gender-specific differences in plasma nitrite and nitrate in adults. Chronic NaNO3 supplementation to healthy young men does not affect renal CA-dependent nitrite excretion or creatinine synthesis and excretion.